Guenat Olivier T, Generelli Silvia, de Rooij Nicolaas F, Koudelka-Hep Milena, Berthiaume François, Yarmush Martin L
Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts 02114, USA. olivier.guenat@ unine.ch
Anal Chem. 2006 Nov 1;78(21):7453-60. doi: 10.1021/ac0609733.
In this study, we present the development and the characterization of a generic platform for cell culture able to monitor extracellular ionic activities (K+, NH4+) for real-time monitoring of cell-based responses, such as necrosis, apoptosis, or differentiation. The platform for cell culture is equipped with an array of 16 silicon nitride micropipet-based ion-selective microelectrodes with a diameter of either 2 or 6 microm. This array is located at the bottom of a 200-microm-wide and 350-microm-deep microwell where the cells are cultured. The characterization of the ion-selective microelectrode arrays in different standard and physiological solutions is presented. Near-Nernstian slopes were obtained for potassium- (58.6 +/- 0.8 mV/pK, n = 15) and ammonium-selective microelectrodes (59.4 +/- 3.9 mV/pNH4, n = 13). The calibration curves were highly reproducible and showed an average drift of 4.4 +/- 2.3 mV/h (n = 10). Long-term behavior and response after immersion in physiological solutions are also presented. The lifetime of the sensors was found to be extremely long with a high recovery rate.
在本研究中,我们展示了一种用于细胞培养的通用平台的开发与特性,该平台能够监测细胞外离子活性(K⁺、NH₄⁺),以实时监测基于细胞的反应,如坏死、凋亡或分化。细胞培养平台配备了由16个基于氮化硅微吸管的离子选择性微电极组成的阵列,其直径为2或6微米。该阵列位于一个宽200微米、深350微米的微孔底部,细胞在此培养。文中展示了离子选择性微电极阵列在不同标准溶液和生理溶液中的特性。钾离子选择性微电极(58.6±0.8 mV/pK,n = 15)和铵离子选择性微电极(59.4±3.9 mV/pNH₄,n = 13)获得了接近能斯特斜率。校准曲线具有高度可重复性,平均漂移为4.4±2.3 mV/h(n = 10)。还展示了在生理溶液中浸泡后的长期行为和响应。发现传感器的寿命极长且恢复率高。